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《Chinese Journal of Hepatobiliary Surgery》 2001-11
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Isolation and establishment of human hepatocellular carcinoma cell clones with various metastatic potentiais

LI Yan TANG Zhaoyou YE Shenglong,Liver Cancer Institute,Zhongshan Hospital,Fudan University,Shanghai 20032,P.R.China  
Objective To establish hepatocellular carcinoma(HCC)cell clones with various me- tastatic potentials from highly metastatic HCC cell line MHCC97.Methods Cell clone culture was conducted on the 25th passage of the source cell line MHCC97 using limited dilution method.Target cell clones were selected by in vivo screening in nude mice.Biological characteristics of the target clones were studied in terms of tumor cell doubling time,karyotyping,in vitro invasiveness,alpha-fe- toprotein secretion and pulmonary metastasis after orthotopic implantation.Results HCC clones with high(MHCC97-H)and low(MHCC97-L)metastatic potentials were established from the source cell line MHCC97.When MHCC97-H was compared with MHCC97-L,the pulmonary metastatic rate was 100%(10/10)versus 40%(4/10),tumor cell doubling time 34.2 hours vs.60.0 hours,and Boyden chamber in vitro invasion assay was 37.5±11.0 cells/field vs.17.74±6.3 cells/field.Karyotyping re- vealed that the main image of chromosomes was 55—57(68%)in MHCC97-H and 57—62(58%)in MHCC97-L.The proportions of cells in G_0—G_1 phase,S phase,and G_2-M phase for MHCC97-H/ MHCC97-L were 56.10%/65.32%, 28.35%/24.96% and 15.64%/9.73%,respectively,as meas- ured by flow cytometry.The serum AFP levels in nude mice in 5 weeks after orthotopic implantation of tumor tissue were 245.8±66.1μg/L for MHCC97-H and 91.2±65.7μg/L for MHCC97-L.Con- clusions The results confirm the heterogeneity of MHCC97.These tow clones of the same genetic background are of great value for further investigation on metastasis of HCC.
【Fund】: 国家重点基础研究发展规划“恶性肿瘤发生与发展的基础性研究”G1998051211 ;; 上海市医学领先专业重大项目:“肝癌转移的分子机制及干预治疗的实验研究”983001
【CateGory Index】: R735.7
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